LOCUS       sequence-210790-        6149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse MeCP2 (R306C) in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6149)
  AUTHORS   Tillotson R, Selfridge J, Koerner MV, Gadalla KKE, Guy J, De Sousa 
            D, Hector RD, Cobb SR, Bird A
  TITLE     Radically truncated MeCP2 rescues Rett syndrome-like neurological 
            defects.
  JOURNAL   Nature. 2017 Oct 19;550(7676):398-401. doi: 10.1038/nature24058. 
            Epub 2017 Oct 11.
  PUBMED    29019980
REFERENCE   2  (bases 1 to 6149)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature24058"; journalName: "Nature"; date: "2017-10-19- 
            19"; volume: "550"; issue: "7676"; pages: "398-401"
COMMENT     Sequence Label: pEGFP-N1_MeCP2(R306C)
FEATURES             Location/Qualifiers
     source          1..6149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1709..1729
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      2084..2093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2090..2092,2093..2095,2096..2809)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2090..2092,2093..2095,2096..2806)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2135..2156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2396..2415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2743..2764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2932..3053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2969..2988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3023..3042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3060..3515)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3197..3218)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3409..3428
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3542..3646
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3648..4005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3648..3839
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3671..3691)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3856..3991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4040..4834
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4040..4834
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4094..4113)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4704..4723
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5022..5041)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5066..5113
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5442..6030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5931..5950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
      601 ctcagatctc gagatggtag ctgggatgtt agggctcagg gaggaaaagt cagaagacca
      661 ggatctccag ggcctcagag acaagccact gaagtttaag aaggcgaaga aagacaagaa
      721 ggaggacaaa gaaggcaagc atgagccact acaaccttca gcccaccatt ctgcagagcc
      781 agcagaggca ggcaaagcag aaacatcaga aagctcaggc tctgccccag cagtgccaga
      841 agcctcggct tcccccaaac agcggcgctc cattatccgt gaccggggac ctatgtatga
      901 tgaccccacc ttgcctgaag gttggacacg aaagcttaaa caaaggaagt ctggccgatc
      961 tgctggaaag tatgatgtat atttgatcaa tccccaggga aaagcttttc gctctaaagt
     1021 agaattgatt gcatactttg aaaaggtggg agacacctcc ttggacccta atgattttga
     1081 cttcacggta actgggagag ggagcccctc caggagagag cagaaaccac ctaagaagcc
     1141 caaatctccc aaagctccag gaactggcag gggtcgggga cgccccaaag ggagcggcac
     1201 tgggagacca aaggcagcag catcagaagg tgttcaggtg aaaagggtcc tggagaagag
     1261 ccctgggaaa cttgttgtca agatgccttt ccaagcatcg cctgggggta agggtgaggg
     1321 aggtggggct accacatctg cccaggtcat ggtgatcaaa cgccctggca gaaagcgaaa
     1381 agctgaagct gacccccagg ccattcctaa gaaacggggt agaaagcctg ggagtgtggt
     1441 ggcagctgct gcagctgagg ccaaaaagaa agccgtgaag gagtcttcca tacggtctgt
     1501 gcatgagact gtgctcccca tcaagaagtg caagacccgg gagacggtca gcatcgaggt
     1561 caaggaagtg gtgaagcccc tgctggtgtc cacccttggt gagaaaagcg ggaagggact
     1621 gaagacctgc aagagccctg ggcgtaaaag caaggagagc agccccaagg ggcgcagcag
     1681 cagtgcctcc tccccaccta agaaggagca ccatcatcac caccatcact cagagtccac
     1741 aaaggccccc atgccactgc tcccatcccc acccccacct gagcctgaga gctctgagga
     1801 ccccatcagc ccccctgagc ctcaggactt gagcagcagc atctgcaaag aagagaagat
     1861 gccccgagga ggctcactgg aaagcgatgg ctgccccaag gagccagcta agactcagcc
     1921 tatggtcgcc accactacca cagttgcaga aaagtacaaa caccgagggg agggagagcg
     1981 caaagacatt gtttcatctt ccatgccaag gccaaacaga gaggagcctg tggacagccg
     2041 gacgcccgtg accgagagag ttagctgtaa ggatccaccg gtcgccacca tggtgagcaa
     2101 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2161 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2221 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2281 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2341 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2401 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2461 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2521 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2581 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2641 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2701 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2761 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     2821 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2881 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2941 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3001 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     3061 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3121 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3181 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3241 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3301 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3361 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3421 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3481 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     3541 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3601 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     3661 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3841 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3901 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3961 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     4021 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4081 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4141 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4201 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4261 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4321 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4381 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4441 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4501 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4561 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4621 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4681 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4741 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4801 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4861 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4921 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4981 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     5041 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     5101 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     5161 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     5221 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     5281 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5341 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5581 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6121 gataaccgta ttaccgccat gcattagtt
//
